Abstract
Abstract The enzyme urate oxidase (UOX) is used as a drug for preventing and treatment of hyperuricemia. This study deals with the statistical optimization of lactose inducible fermentation for production of soluble recombinant Aspergillus flavus UOX. 10 independent variables were investigated by Plackett–Burman design (PBD), and the most significant factors were further optimized by central composite design (CCD). PBD results indicated that glycerol, yeast extract, tryptone, and lactose affected UOX activity significantly. The CCD results showed that the maximum enzyme activity (19.34 U/ml) and biomass concentration (44.0 g/L) can be achieved under the optimum conditions of glycerol 0.87 g/L, yeast extract 9.11 g/L, tryptone 10.29 g/L, K2HPO4 1.81 g/L, and lactose 12.79 g/L. When the same induction strategy was tested at shake flask, 19.34 U/mL of UOX activity was obtained, which was 12.5 folds higher than IPTG induction protocol. Furthermore, the lower total cost (€0.7 vs. €13.5) was additional feature that confirmed the suitability of the lactose induction method. Collectively, our results showed that design of experiment methodology can be applied as a suitable tool for improved production of UOX enzyme using lactose as the inducer.
Highlights
Uric acid is the final product of purine metabolism in hominoids
Plackett–Burman design (PBD) was applied to screen the important variables that significantly affect the production of urate oxidase (UOX) enzyme
Large positive or negative coefficients indicate that these factors including A, B, C, K and D play an important role in UOX production, while J, H, E and G indicates negative effects
Summary
Uric acid is the final product of purine metabolism in hominoids. Because of low solubility of uric acid, various species have specific enzymes to change it into more soluble substances such as allantoin (Hayashi et al, 2000). Because of the industrial importance of UOX as a drug enzyme, various studies have focused on the production optimization, and cost effective bioprocessing. Uox gene from A. flavus was expressed in Escherichia coli BL21 (DE3) using lactose inducible expression system, and statistical methods were applied to increase the production yield.
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